Trophic transfer increases the exposure to microplastics in littoral predators.

Predators in aquatic environments can be exposed to microplastics (MPs) directly through water and indirectly through prey. Laboratory experiments were conducted to study the potential of MP trophic transfer in Baltic Sea littoral food chains of different lengths. The longest studied food chain had three trophic levels: zooplankton, chameleon shrimp (Praunus flexuosus) and rockpool prawn (Palaemon elegans). 10 µm fluorescence microspheres were used as tracer MP particles and MP ingestion was verified with epifluorescence microscopy. Transfer of MPs occurred up to both second and third trophic level. The number of ingested microspheres in both chameleon shrimp and rockpool prawn was higher when the animals were exposed through pre-exposed prey in comparison to direct exposure through the water. The results show that trophic transfer may be an important pathway of and increase the microplastic exposure for some animals at higher trophic levels in highly polluted areas.

Testing citizen science as a tool for monitoring surface water microplastics.

The use of citizen science in the collection of surface water marine microplastics (MP) samples with manta trawl was tested in the Baltic Sea, where the collection of surface water samples is often hampered by environmental conditions. Sampling was carried out at 7 locations around the Baltic Sea with a custom-made manta trawl which was operated onboard a sailing boat. The total concentrations of ≥ 0.3 mm MP in the samples ranged from 0.45 to 1.98 MP m-3. Based on the results and experiences from this study, citizen science could be introduced into the toolbox of monitoring large MP. When the common basic constraints of surface water sampling within a regional sea are defined and agreed upon, citizen science could be used for strengthening the power of assessments on the state of the marine environment by increasing the spatial coverage of the monitored area.

Exposure to leachates from post-consumer plastic and recycled rubber causes stress responses and mortality in a copepod Limnocalanus macrurus.

Effects of household post-consumer plastics and tyre rubber on a Baltic Sea copepod Limnocalanus macrurus were assessed. Fragments of commercial recycled low-density polyethylene vegetable bags and rubber originating from recycled car tyres were incubated in seawater, and the copepods were exposed to the filtrate of the water. L. macrurus experienced erratic swimming behaviour and increased mortality in the filtrate of unwashed vegetable bags, containing elevated concentrations of alcohols, organic acids and copper. Responses of the antioxidant defence system (ADS) were recorded in copepods exposed to rubber treatments containing high concentrations of zinc. Significant responses in the ADS enzymes indicate that reactive oxygen species (ROS) formation was exceeding the detoxification capacity of the ADS which may further lead to prolonged state of oxidative stress. Observed effects of exposure on the biochemical level coincide with impaired swimming activity of the copepods, indicating possible irreversible cellular responses leading to behavioural changes and mortality.

Microplastic ingestion by small coastal fish in the northern Baltic Sea, Finland.

Microplastic (MP) ingestion by four species of small coastal fish from the northern Baltic Sea was investigated. The digestive tract contents of 424 specimens, caught across eight sampling sites along the Finnish coastline were analysed for the occurrence of MP ingestion. MP were found in 38 fish individuals (9% of sampled fish). Specimens from the urban area of Helsinki displayed the highest prevalence of ingested plastics (27.5%). No relationship was found between the size or species of the fish and the presence of ingested MP particles nor the amount of MP in seawater. The comparison to a previous study conducted using the same research methods indicates that the ingestion of MP is more common in coastal fish than in offshore fish in the northern Baltic Sea.

Validation of an imaging FTIR spectroscopic method for analyzing microplastics ingestion by Finnish lake fish (Perca fluviatilis and Coregonus albula).

Despite the ubiquitousness of microplastics, knowledge on the exposure of freshwater fish to microplastics is still limited. Moreover, no standard methods are available for analyzing microplastics, and the quality of methods used for the quantification of ingested microplastics in fish should be improved. In this study, we studied microplastic ingestion of common wild freshwater fish species, perch (Perca fluviatilis) and vendace (Coregonus albula). Further, our aim was to develop and validate imaging Fourier-transform infrared spectroscopic method for the quantification of ingested microplastics. For this purpose, enzymatically digested samples were measured with focal plane array (FPA) based infrared microscope. Data was analyzed with siMPle software, which provides counts, mass estimations, sizes, and materials for the measured particles. Method validation was conducted with ten procedural blanks and recovery tests, resulting in 75% and 77% recovery rates for pretreatment and infrared imaging, respectively. Pretreatment caused contamination principally by small <100 µm microplastics. The results showed that 17% of perch and 25% of vendace had ingested plastic. Most of the fish contained little or no plastics, while some individuals contained high numbers of small particles or alternatively few large particles. Perch from one sampling site out of five had ingested microplastics, but vendace from all sampling sites had ingested microplastics. The microplastics found from fish were mostly small: 81% had particle size between 20 and 100 µm, and most of them were polyethylene, polypropylene, and polyethylene terephthalate. In conclusion, the implemented method revealed low numbers of ingested microplastics on average but needs further development for routine monitoring of small microplastics.

Microplastics accumulate to thin layers in the stratified Baltic Sea.

In the Baltic Sea, water is stratified due to differences in density and salinity. The stratification prevents water from mixing, which could affect sinking rates of microplastics in the sea. We studied the accumulation of microplastics to halocline and thermocline. We sampled water with a 100 µm plankton net from vertical transects between halo- and thermocline, and a 30 L water sampler from the end of halocline and the beginning of thermocline. Thereafter, microplastics in the whole sample volumes were analyzed with imaging Fourier transform infrared spectroscopy (FTIR). The plankton net results showed that water column between halo- and thermoclines contained on average 0.92 ± 0.61 MP m-3 (237 ± 277 ng/m-3; mean ± SD), whereas the 30 L samples from the end of halocline and the beginning of thermocline contained 0.44 ± 0.52 MP L-1 (106 ± 209 ng L-1). Hence, microplastics are likely to accumulate to thin layers in the halocline and thermocline. The vast majority of the found microplastics were polyethylene, polypropylene and polyethylene terephthalate, which are common plastic types. We did not observe any trend between the density of microplastics and the sampling depth, probably because biofilm formation affected the sinking rates of the particles. Our results indicate the need to sample deeper water layers in addition to surface waters at least in the stratified water bodies to obtain a comprehensive overview of the abundance of microplastics in the aquatic environment.

Polycyclic aromatic hydrocarbon sorption and bacterial community composition of biodegradable and conventional plastics incubated in coastal sediments.

Resistant to degradation, plastic litter poses a long-term threat to marine ecosystems. Biodegradable materials have been developed to replace conventional plastics, but little is known of their impacts and degradation in marine environments. A 14-week laboratory experiment was conducted to investigate the sorption of polycyclic aromatic hydrocarbons (PAHs) to conventional (polystyrene PS and polyamide PA) and bio-based, biodegradable plastic films (cellulose acetate CA and poly-L-lactic acid PLLA), and to examine the composition of bacterial communities colonizing these materials. Mesoplastics (1 cm2) of these materials were incubated in sediment and seawater collected from two sites in the Gulf of Finland, on the coast of the highly urbanized area of Helsinki, Finland. PS sorbed more PAHs than did the other plastic types at both sites, and the concentration of PAHs was consistently and considerably smaller in plastics than in the sediment. In general, the plastic bacterial biofilms resembled those in the surrounding media (water and/or sediment). However, in the sediment incubations, the community composition on CA diverged from that of the other three plastic types and was enriched with Bacteroidia and potentially cellulolytic Spirochaetia at both sites. The results indicate that certain biodegradable plastics, such as CA, may harbour potential bioplastic-degrading communities and that PAH sorption capacity varies between polymer types. Since biodegradable plastics are presented as replacements for conventional plastics in applications with risk of ending up in the marine environment, the results highlight the need to carefully examine the environmental behaviour of each biodegradable plastic type before they are extensively introduced to the market.

Microplastic concentrations, size distribution, and polymer types in the surface waters of a northern European lake.

We examined microplastic concentrations, size distributions, and polymer types in surface waters of a northern European dimictic lake. Two sampling methods, a pump sieving water onto filters with different pore sizes (20, 100, and 300 µm) and a common manta trawl (333 µm), were utilized to sample surface water from 12 sites at the vicinity of potential sources for microplastic emissions. The number and polymer types of microplastics in the samples were determined with optical microscopy and µFTIR spectroscopy. The average concentrations were 0.27 ± 0.18 (mean ± SD) microplastics/m3 in manta trawled samples and 1.8 ± 2.3 (>300 µm), 12 ± 17 (100-300 µm) and 155 ± 73 (20-100 µm) microplastics/m3 in pump filtered samples. The majority (64%) of the identified microplastics (n = 168) were fibers, and the rest were fragments. Materials were identified as polymers commonly used in consumer products, such as polyethylene, polypropylene, and polyethylene terephthalate. Microplastic concentrations were high near the discharge pipe of a wastewater treatment plant, harbors, and snow dumping site. PRACTITIONER POINTS: Samples were taken with a manta trawl (333 µm) and a pump filtration system (300/100/20 µm) With pump filtration, small 20-300 µm particles were more common than >300 µm particles The average concentration of manta trawled samples was 0.27 ± 0.18 (mean ± SD) microplastics/m3 FTIR analysis revealed PE, PP, PET, and PAN to be the most common polymers.

Seafloor sediments as microplastic sinks in the northern Baltic Sea - Negligible upward transport of buried microplastics by bioturbation.

Microplastics (MPs) are ubiquitous in the marine environment. High concentrations of MPs are found from seafloor sediments, which have been proposed to act as their final sinks. Because bioturbation is an important process affecting the burial of MPs, a mesocosm experiment was established to study whether sediment infauna may also promote MP return to the sediment surface. Thin layers of frozen sediment containing an environmentally realistic concentration (<1300 MPs per kg of dry sediment) of MP fragments in two size classes (>500 µm and 100-300 µm) were added to depths of 2 cm and 5 cm in the experimental cylinders filled with sediment. The displacement of these MPs, made of acrylonitrile butadiene styrene (ABS), by a community of common benthic invertebrates in the northern Baltic Sea (clam Limecola balthica, polychaete Marenzelleria spp., gammarid Monoporeia affinis) was studied in a 10-week experiment. After the experiment, the MPs were extracted from each sediment layer and the animals were examined for MP ingestion. The results indicated that the transportation of MPs to the sediment surface by bioturbation was negligible. Thus, in the Baltic Sea, the seafloor may act as a sink for once sedimented MPs, reducing simultaneously the MP exposure of the macrofauna feeding on the sediment surface.

Application of an enzyme digestion method reveals microlitter in Mytilus trossulus at a wastewater discharge area.

The ingestion of microlitter by blue mussels (450) was studied at a wastewater recipient area in the Baltic Sea. The mussel soft tissues were digested using enzymatic detergents and the detected litter particles characterized with FT-IR imaging spectroscopy. Microlitter concentration in seawater and WWTP effluent were also measured. Microlitter was found in 66% of the mussels. Mussels from the WWTP recipient had higher microlitter content compared to those collected at the reference site. Plastics made up 8% of all the analysed microlitter particles. The dominating litter types were fibres (~90% of all microlitter), 42% of which were cotton, 17% linen, 17% viscose and 4% polyester. The risk of airborne contamination during laboratory work was lowered when mussels were digested with their shells on instead of dissecting them first. The approach was found applicable and gentle to both non-synthetic and synthetic materials including fragile fibres.

Effective and easy to use extraction method shows low numbers of microplastics in offshore planktivorous fish from the northern Baltic Sea.

Although the presence of microplastics in marine biota has been widely recorded, extraction methods, method validation and approaches to monitoring are not standardized. In this study a method for microplastic extraction from fish guts based on a chemical alkaline digestion is presented. The average particle retrieval rate from spiked fish guts, used for method validation, was 84%. The weight and shape of the test particles (PET, PC, HD-PE) were also analysed with no noticeable changes in any particle shapes and only minor weight change in PET (2.63%). Microplastics were found in 1.8% of herrings (n=164) and in 0.9% of sprat (n=154). None of the three-spined sticklebacks (n=355) contained microplastic particles.

Solutions to microplastic pollution - Removal of microplastics from wastewater effluent with advanced wastewater treatment technologies.

Conventional wastewater treatment with primary and secondary treatment processes efficiently remove microplastics (MPs) from the wastewater. Despite the efficient removal, final effluents can act as entrance route of MPs, given the large volumes constantly discharged into the aquatic environments. This study investigated the removal of MPs from effluent in four different municipal wastewater treatment plants utilizing different advanced final-stage treatment technologies. The study included membrane bioreactor treating primary effluent and different tertiary treatment technologies (discfilter, rapid sand filtration and dissolved air flotation) treating secondary effluent. The MBR removed 99.9% of MPs during the treatment (from 6.9 to 0.005 MP L-1), rapid sand filter 97% (from 0.7 to 0.02 MP L-1), dissolved air flotation 95% (from 2.0 to 0.1 MP L-1) and discfilter 40-98.5% (from 0.5 - 2.0 to 0.03-0.3 MP L-1) of the MPs during the treatment. Our study shows that with advanced final-stage wastewater treatment technologies WWTPs can substantially reduce the MP pollution discharged from wastewater treatment plants into the aquatic environments.

Bioturbation transports secondary microplastics to deeper layers in soft marine sediments of the northern Baltic Sea.

Microplastics (MPs) are observed to be present on the seafloor ranging from coastal areas to deep seas. Because bioturbation alters the distribution of natural particles on inhabited soft bottoms, a mesocosm experiment with common benthic invertebrates was conducted to study their effect on the distribution of secondary MPs (different-sized pieces of fishing line<1mm). During the study period of three weeks, the benthic community increased MP concentration in the depth of 1.7-5.1cm in the sediment. The experiment revealed a clear vertical gradient in MP distribution with their abundance being highest in the uppermost parts of the sediment and decreasing with depth. The Baltic clam Macoma balthica was the only study animal that ingested MPs. This study highlights the need to further examine the vertical distribution of MPs in natural sediments to reliably assess their abundance on the seafloor as well as their potential impacts on benthic communities.

How well is microlitter purified from wastewater? - A detailed study on the stepwise removal of microlitter in a tertiary level wastewater treatment plant.

Wastewater treatment plants (WWTPs) can offer a solution to reduce the point source input of microlitter and microplastics into the environment. To evaluate the contributing processes for microlitter removal, the removal of microlitter from wastewater during different treatment steps of mechanical, chemical and biological treatment (activated sludge) and biologically active filter (BAF) in a large (population equivalent 800 000) advanced WWTP was examined. Most of the microlitter was removed already during the pre-treatment and activated sludge treatment further decreased the microlitter concentration. The overall retention capacity of studied WWTP was over 99% and was achieved after secondary treatment. However, despite of the high removal performance, even an advanced WWTP may constitute a considerable source of microlitter and microplastics into the aquatic environment given the large volumes of effluent discharged constantly. The microlitter content of excess sludge, dried sludge and reject water were also examined. According to the balance analyses, approximately 20% of the microlitter removed from the process is recycled back with the reject water, whereas 80% of the microlitter is contained in the dried sludge. The study also looked at easy microlitter sampling protocol with automated composite samplers for possible future monitoring purposes.

Distribution and abundance of surface water microlitter in the Baltic Sea: A comparison of two sampling methods.

Two methods for marine microlitter sampling were compared in the Gulf of Finland, northern Baltic Sea: manta trawl (333µm) and a submersible pump (300 or 100µm). Concentrations of microlitter (microplastics, combustion particles, non-synthetic fibres) in the samples collected with both methods and filter sizes remained <10particlesm(-3). The pump with 100µm filter gave higher microlitter concentrations compared to manta trawl or pump with 300µm filter. Manta sampling covers larger areas, but is potentially subjected to contamination during sample processing and does not give precise volumetric values. Using a submerged pump allows method controls, use of different filter sizes and gives exact volumetric measures. Both devices need relatively calm weather for operation. The choice of the method in general depends on the aim of the study. For monitoring environmentally relevant size fractions of microlitter the use of 100µm or smaller mesh size is recommended for the Baltic Sea.

The Genetic Diversity of Mesodinium and Associated Cryptophytes.

Ciliates from the genus Mesodinium are globally distributed in marine and freshwater ecosystems and may possess either heterotrophic or mixotrophic nutritional modes. Members of the Mesodinium major/rubrum species complex photosynthesize by sequestering and maintaining organelles from cryptophyte prey, and under certain conditions form periodic or recurrent blooms (= red tides). Here, we present an analysis of the genetic diversity of Mesodinium and cryptophyte populations from 10 environmental samples (eight globally dispersed habitats including five Mesodinium blooms), using group-specific primers for Mesodinium partial 18S, ITS, and partial 28S rRNA genes as well as cryptophyte large subunit RuBisCO genes (rbcL). In addition, 22 new cryptophyte and four new M. rubrum cultures were used to extract DNA and sequence rbcL and 18S-ITS-28S genes, respectively, in order to provide a stronger phylogenetic context for our environmental sequences. Bloom samples were analyzed from coastal Brazil, Chile, two Northeastern locations in the United States, and the Pribilof Islands within the Bering Sea. Additionally, samples were also analyzed from the Baltic and Barents Seas and coastal California under non-bloom conditions. Most blooms were dominated by a single Mesodinium genotype, with coastal Brazil and Chile blooms composed of M. major and the Eastern USA blooms dominated by M. rubrum variant B. Sequences from all four blooms were dominated by Teleaulax amphioxeia-like cryptophytes. Non-bloom communities revealed more diverse assemblages of Mesodinium spp., including heterotrophic species and the mixotrophic Mesodinium chamaeleon. Similarly, cryptophyte diversity was also higher in non-bloom samples. Our results confirm that Mesodinium blooms may be caused by M. major, as well as multiple variants of M. rubrum, and further implicate T. amphioxeia as the key cryptophyte species linked to these phenomena in temperate and subtropical regions.

Feeding type affects microplastic ingestion in a coastal invertebrate community.

Marine litter is one of the problems marine ecosystems face at present, coastal habitats and food webs being the most vulnerable as they are closest to the sources of litter. A range of animals (bivalves, free swimming crustaceans and benthic, deposit-feeding animals), of a coastal community of the northern Baltic Sea were exposed to relatively low concentrations of 10 µm microbeads. The experiment was carried out as a small scale mesocosm study to mimic natural habitat. The beads were ingested by all animals in all experimental concentrations (5, 50 and 250 beads mL(-1)). Bivalves (Mytilus trossulus, Macoma balthica) contained significantly higher amounts of beads compared with the other groups. Free-swimming crustaceans ingested more beads compared with the benthic animals that were feeding only on the sediment surface. Ingestion of the beads was concluded to be the result of particle concentration, feeding mode and the encounter rate in a patchy environment.

Do wastewater treatment plants act as a potential point source of microplastics? Preliminary study in the coastal Gulf of Finland, Baltic Sea.

This study on the removal of microplastics during different wastewater treatment unit processes was carried out at Viikinmäki wastewater treatment plant (WWTP). The amount of microplastics in the influent was high, but it decreased significantly during the treatment process. The major part of the fibres were removed already in primary sedimentation whereas synthetic particles settled mostly in secondary sedimentation. Biological filtration further improved the removal. A proportion of the microplastic load also passed the treatment and was found in the effluent, entering the receiving water body. After the treatment process, an average of 4.9 (±1.4) fibres and 8.6 (±2.5) particles were found per litre of wastewater. The total textile fibre concentration in the samples collected from the surface waters in the Helsinki archipelago varied between 0.01 and 0.65 fibres per litre, while the synthetic particle concentration varied between 0.5 and 9.4 particles per litre. The average fibre concentration was 25 times higher and the particle concentration was three times higher in the effluent compared to the receiving body of water. This indicates that WWTPs may operate as a route for microplastics entering the sea.

Rhinomonas nottbecki n. sp. (cryptomonadales) and molecular phylogeny of the family Pyrenomonadaceae.

The cryptomonad Rhinomonas nottbecki n. sp., isolated from the Baltic Sea, is described from live and fixed cells studied by light, scanning, and transmission electron microscopy together with sequences of the partial nucleus- and nucleomorph-encoded 18S rRNA genes as well as the nucleus-encoded ITS1, 5.8S, ITS2, and the 5'-end of the 28S rRNA gene regions. The sequence analyses include comparison with 43 strains from the family Pyrenomonadaceae. Rhinomonas nottbecki cells are dorsoventrally flattened, obloid in shape; 10.0-17.2 µm long, 5.5-8.1 µm thick, and 4.4-8.8 µm wide. The inner periplast has roughly hexagonal plates. Rhinomonas nottbecki cells resemble those of Rhinomonas reticulata, but the nucleomorph 18S rRNA gene of R. nottbecki differs by 2% from that of R. reticulata, while the ITS region by 11%. The intraspecific variability in the ITS region of R. nottbecki is 5%. In addition, the predicted ITS2 secondary structures are different in R. nottbecki and R. reticulata. The family Pyrenomonadaceae includes three clades: Clade A, Clade B, and Clade C. All Rhinomonas sequences branched within the Clade C, while the genus Rhodomonas is paraphyletic. The analyses suggest that the genus Storeatula is an alternating morphotype of the genera Rhinomonas and Rhodomonas and that the family Pyrenomonadaceae includes some species that were described multiple times, as well as novel species.

Ingestion and transfer of microplastics in the planktonic food web.

Experiments were carried out with different Baltic Sea zooplankton taxa to scan their potential to ingest plastics. Mysid shrimps, copepods, cladocerans, rotifers, polychaete larvae and ciliates were exposed to 10 µm fluorescent polystyrene microspheres. These experiments showed ingestion of microspheres in all taxa studied. The highest percentage of individuals with ingested spheres was found in pelagic polychaete larvae, Marenzelleria spp. Experiments with the copepod Eurytemora affinis and the mysid shrimp Neomysis integer showed egestion of microspheres within 12 h. Food web transfer experiments were done by offering zooplankton labelled with ingested microspheres to mysid shrimps. Microscopy observations of mysid intestine showed the presence of zooplankton prey and microspheres after 3 h incubation. This study shows for the first time the potential of plastic microparticle transfer via planktonic organisms from one trophic level (mesozooplankton) to a higher level (macrozooplankton). The impacts of plastic transfer and possible accumulation in the food web need further investigations.